Factor VII and related polypeptides have been demonstrated to be useful therapeutic agents. Accordingly, there is an increasing need for formulations comprising these proteins that are pharmaceutically acceptable and exhibit a uniform and predetermined clinical efficacy.
The overall industrial-scale process for the purification of a Factor VII polypeptide drug substance may suffer from the drawback that an initial drug substance considerable amount of product-related (Factor VII-related) impurities (such as impurities contained in late eluting peaks (including, e.g., glyco-variants with differing levels of N-linked glycosylation (“des-N-glycan forms”)), oxidized forms, proteolytically degraded forms (heavy-chain cleaved forms), or aggregates having a higher molecular weight than the Factor VII polypeptide of interest)).
The article “Amino Acid Sequence and Posttranslational Modifications of Human Factor VIIa from Plasma and Transfected Baby Hamster Kidney Cells”, Biochemistry, 1988 Oct. 4; 27(20):7785-93, reports that some heavy chain degradation products co-purify with intact activated Factor VII.
The article “FVIIa Derivatives Obtained by Autolytic and Controlled Cathepsin G Mediated Cleavage”, FEBS Lett. 1993 Feb. 15; 317(3):245-9, states that heavy chain cleaved forms of Factor VII cannot be isolated from Factor VII under non-denaturing conditions.
U.S. Pat. No. 6,777,390 (Baxter) concerns purification of factor VII from cryosupernatant by anion exchange and subsequent hydrophobic chromatography on Phenyl-Sepharose.
A problem that also or alternatively may be associated with the preparation of such polypeptides is that the polypeptide may not have a uniform glycosylation pattern.
There remains a need for Factor VII Polypeptide preparations, having reduced amounts of product-related impurities (e.g., Factor VII Polypeptide drug substances that are free or at least substantially free of late eluting peaks) and/or a uniform glycosylation pattern, and methods for making such preparations, particularly in industrial scale quantities. The invention described herein provides new methods for preparing such compositions. This and other advantages and further aspects and features of the invention will be apparent from the description of the invention provided here.